OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Eph family tyrosine kinase receptors consist of two groups: EphA group members respond to ephrin-A, which is anchored to the cell membrane by glycosylphosphatidylinositol, whereas EphB group members respond to ephrin-B, containing a transmembrane domain. Recruitment of CrkII and activation of Rho signalling are responsible for EphA3-mediated cell rounding, blebbing and de-adhesion. This response is activated by ephrin-A5-mediated receptor clustering and EphA3 tyrosine kinase activity.
Family:
Kinase
Sub Family:
RTK
Class:
Protein Tyrosine
Protein Name:
EphA3
Uniprot Number:
P29320
Protein Aliases:
EPH-like kinase 4|EK4|hEK4|HEK|Human embryo kinase|Tyrosine-protein kinase TYRO4|Tyrosine-protein kinase receptor ETK1|Eph-like tyrosine kinase 1
Gene Name:
EPHA3
Gene ID:
2042
Gene Aliases:
HEK|HEK4
Accession Number:
NM_005233
Organism:
Human
Construct Details:
578-end recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometric
Measured Response:
Scintillation
Testing Information
Procedure Summary:
EphA3 (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 0.1 mg/mL poly (Glu,Tyr) 4:1, 10 mM MgAcetate and [gamma-33P]-ATP (specific activity and concentration as required). The reaction is initiated by the addition of the Mg/ATP mix. After incubation for 40 minutes at room temperature, the reaction is stopped by the addition of phosphoric acid to a concentration of 0.5%. An aliquot of the reaction is then spotted onto a filter and washed four times for 4 minutes in 0.425% phosphoric acid and once in ethanol prior to drying and scintillation counting.
Substrate:
0.1 mg/mL poly (Glu,Tyr)
Tracer:
33P
ATP Concentration:
[ATP]= 120µM (Km= 119µM )
Incubation:
40 min at Room temperature
Control Inhibitor:
Staurosporine
Test Sample Requirements:
Please provide compounds dissolved in 100% DMSO (PBS or HBSS prohibited).
Stock solution should be ≥50× the final assay concentration (a 10 mM stock is also acceptable).
Required volume:
– <60 kinase assays: 120 µL of 50× stock
– ≥60 assays: 420 µL of 50× stock
If you supply 10 mM stocks or powder (pre-weighed), we will apply the appropriate conversion.
For batches of ~50 compounds supplied as solids, the TAT will be extended.
Minimum Order Quantity:
2
Turnaround Time
Standard:
6 Business Days
Service Scheduling Note:
Assay is run on a weekly schedule beginning on Friday. Compounds and purchase order must be received on or before 13:00 CET Thursday for the order to be included on the service run that begins on Friday of the same week.
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Oncology/Immuno-Oncology
Adverse / Beneficial Action:
Inhibition of EPHA3 may inhibit tumor progression as well as promote axonal regeneration following spinal cord injury.
Additional Information
Brand:
LeadHunter
Testing Location:
KP - France - Celle L'Evescault
RELATED SOLUTIONS
EphA3 Human RTK Kinase Enzymatic Radiometric [10 uM ATP] KinaseProfiler LeadHunter Assay - FR
Item: 14-644KP10
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